Abstract Coffee crop in Brazil is typically grown as a monoculture. However, we hypothesized that agroforestry system is favorable association for arbuscular mycorrhizal fungi (AMF), affecting its community structure and potentially impacting crop productivity and agroecosystem health. This study evaluated how the microclimate, soil depth, macauba field spacing and distance between coffee plants and palms affect the structure of the AMF community. The structure of the AMF community was influenced by the soil depth, microclimate features, soil moisture, maximum air temperature, and photosynthetically active radiation (PAR). The distance at which coffee-macauba influences ecological diversity indices of AMF, and higher diversity are related to the proximity between plants. AMF diversity (Richness and Shannon) in the agroforestry system exceeded that observed in the full-sun coffee in the 0-20 soil depth layer. Our results showed that the microclimate, soil depth, plant density, and distance between coffee from macauba affected the AMF community structure.

ABSTRACT Restoration of degraded sites by using native plants like Plathymenia reticulata Benth. and Melanoxylon brauna Schot. is advisable. However, seedlings of both species, when raised on commercial substrates, may present low survival. This study aimed to evaluate the growth of seedlings under inoculation with arbuscular mycorrhizal fungi (AMF). The seedlings were raised on soil sampled from near an adult plant of the same species. The P. reticulata and M. brauna seedlings were grown with or without the inoculation of a mix of three species of AMF (Rhizophagus clarus, Claroideoglomus etunicatum, and Gigaspora albida), and five doses of P (0, 50, 150, 300, and 450 mg dm-3 of P). All seedlings were inoculated with Bradyrhizobium sp., isolated from each species. After 90 days, P. reticulata inoculated with AMF showed higher growth and nutrient content than those uninoculated, and the maximum plant growth was obtained when received 160 to 280 mg dm-3 of P. On the other hand, no effects of AMF inoculation or phosphate fertilization were observed on M. brauna. For both species, no effect of inoculation was observed on mycorrhizal colonization, and M. brauna, unlike P. reticulata presented a considerable number of nodules. We conclude that inoculation with AMF and P fertilization improves the growth of P. reticulata seedlings but does not promote the growth of M. brauna; presenting the necessity to investigate each species.

ABSTRACT The production of inoculum from arbuscular mycorrhizal fungi (AMF) at a large scale and low cost is essential for establishing methods to assist in producing pineapple plantlets with high nutritional and phytosanitary quality. However, this objective is difficult to accomplish because of the biotrophic nature of these fungi. The on-farm multiplication method for AMF inoculum presents a good alternative to supply the demand for the production of glomerospores. This study aimed to multiply and evaluate AMF inoculum originating from isolated species (including Rhizophagus clarus, Claroideoglomus etunicatum) versus native AMF from pineapple and coffee plantations multiplied by the on-farm method on the colonization in pineapple plantlets. Initially, inocula of R. clarus, C. etunicatum, and native AMF (pineapple and coffee) were multiplied by the on-farm method in Sorghum bicolor. After four months, the number of AMF spores and the percentage of viable spores at the layers of 0.00-0.05 and 0.05-0.10 m were evaluated. There were no differences in spore numbers in relation to the source of the inoculum (R. clarus, C. etunicatum, pineapple, and coffee) and evaluated layers, with an average number of 605 spores per 100 cm3 of soil. The percentage of viable spores was greater at the layer of 0.00-0.05 m (76.32 %) compared to the layer of 0.05-0.10 m (72.05 %), regardless of the inoculum source. The viability of the inoculum obtained from C. etunicatum was higher than that from the coffee crop (77.93 and 68.06 %, respectively). Subsequently, the spores were inoculated in pineapple plantlets to assess the rate of colonization. Pineapple plantlets inoculated with AMF had an average of colonization of 18 and 67.73 % after 50 and 180 days cultivation, respectively, with no significant difference being detected between treatments. Therefore, on-farm inoculum production was effective at multiplying the AMF of both isolates of R. clarus and C. etunicatum, as well as for commercial crops (pineapple and coffee), with spores having high viability. Arbuscular mycorrhizal fungi colonized pineapple plantlets independently of the inoculum utilized and favored its growth.

ABSTRACT The multi-enzyme complex (crude extract) of white rot fungi Pleurotus ostreatus, Pleurotus eryngii, Trametes versicolor, Pycnosporus sanguineus and Phanerochaete chrysosporium were characterized, evaluated in the hydrolysis of pretreated pulps of sorghum straw and compared efficiency with commercial enzyme. Most fungi complexes had better hydrolysis rates compared with purified commercial enzyme.

ABSTRACT Symbiosis between orchid seeds and mycorrhizal fungi has been reported to be a determining factor in the success of germination and protocorm development in vitro. The aim of this study was to isolate and identify by molecular analysis the mycorrhizal fungus associated with Cyrtopodium paludicolum, and to evaluate its efficiency in facilitating seed germination and development. Germination experiments were carried out using a fungus isolated from C. paludicolum (CH01) and Epidendrum secundum (M65), which has been successfully used a number of times in symbiotic germination. The experiments were conducted in a completely randomized design with treatments of CH01, M65 as well as under asymbiotic conditions. The mycobiont CH01 was successfully isolated from Cyrtopodium paludicolum and identified as Tulasnella sp. Treatments with both fungi reached a higher germination percentage than under asymbiotic conditions, indicating no specificity in the relationship between Cyrtopodium paludicolum and the fungi. The results presented have the potential to advance research into the propagation and conservation of C. paludicolum, a native of the Cerrado biome.

RESUMO O fungo Pochonia chlamydosporia é um dos mais estudados agentes de controle biológico de fitonematoides. Neste estudo foi demostrado que os isolados Pc-3, Pc-10 e Pc-19 do fungo promovem o crescimento de plântulas de tomate e alface. O isolado Pc-19 colonizou o rizoplano de mudas de tomate em apenas 15 dias e produziu grande quantidade de clamidósporos. O isolado foi capaz de utilizar celulose como fonte de carbono, tanto quanto glicose e sacarose. Em estudos realizados por microscopia eletrônica de varredura (MEV), foi possível observar que hifas do isolado Pc-10 penetraram as células epidérmicas das raízes de tomateiro. Os três isolados de P. chlamydosporia promovem o crescimento de tomateiro e alface.

ABSTRACT The fungus Pochonia chlamydosporia is one of the most studied biological agents used to control plant-parasitic nematodes. This study found that the isolates Pc-3, Pc-10 and Pc-19 of this fungus promote the growth of tomato and lettuce seedlings. The isolate Pc-19 colonized the rhizoplane of tomato seedlings in only 15 days and produced a large quantity of chlamydospores. This isolate was able to use cellulose as a carbon source, in addition to glucose and sucrose. Scanning electron microscopy (SEM) revealed that hyphae of the P. chlamydosporia isolate Pc-10 penetrated the epidermal cells of the tomato roots. These three P. chlamydosporia isolates promote the growth of tomato and lettuce.

Cyrtopodium glutiniferum is an endemic orchid of Brazil with potential medicinal and ornamental applications. As mycorrhizal fungi are essential for the initiation of the orchid life cycle, the aim of this study was to determine the strains of mycorrhizal fungi suitable for seed germination and protocorm development of C. glutiniferum and to characterize the symbiotic development of protocorms. Seeds of C. glutiniferum were inoculated with nine mycorrhizal fungi, Epulorhiza spp., Ceratorhiza spp., Rhizoctonia sp., originally isolated from Brazilian neotropical orchids. Only Epulorhiza isolates promoted seed germination and protocorm development. Three Epulorhiza isolates (M1, M6 = E. epiphytica, M20 = Epulorhiza sp.) promoted protocorm development until leaf production at 63 days. The protocorms are comprised of parenchyma cells delimited by a unistratified epidermis; the parenchyma cells of the upper part of the protocorms are smaller than those located more towards the base. Intact and digested pelotons were observed inside of protocorms implying that the seedlings were capable of mycotrophy. Additionally, the development of a bud primordium only occurred after colonization by fungus. This study suggests that C. glutiniferum has a preference for strains of Epulorhiza and that fungus digestion is essential to protocorm development.

Mycorrhizal associations can be considered required for orchids, which depend on the fungi for germination and establishment in natural conditions. Knowledge of the mycorrhizal fungi is important for programs aimed at the reintroduction, conservation and management of orchid species. The objective of this study was the molecular characterization of Rhizoctonia-like fungi from orchids in the Quadrilátero Ferrífero ("Iron Quadrangle") and Zona da Mata ("Forest Zone") regions of the state of Minas Gerais, Brazil. The affinities of these fungi were studied by comparing the rRNA internal transcribed spacer region with that of other isolates and sequences in GenBank. Three isolates had an affinity for Epulorhiza repens, and one was the holotypeof E. epiphytica.

Culturas monoxênicas de fungos micorrízicos arbusculares (FMAs) em raízes transformadas têm sido utilizadas para melhor compreensão desses fungos, porém, poucas espécies foram estabelecidas in vitro com sucesso. O objetivo do trabalho foi estabelecer culturas monoxênicas de Gigaspora decipiens e Glomus clarum e verificar o efeito da temperatura e pH na produção de esporos. As culturas foram obtidas de esporos desinfestados e germinados em Agar-água. Após a germinação foram transferidos para placas de Petri contendo raízes de cenoura transformadas, em meio mínimo. Após 4 a 6 dias formou-se novos esporos e estruturas típicas. O efeito da temperatura (22, 25, 28 e 32ºC) e pH (4,0; 4,5; 5,5 e 6,5) na produção de esporos foram analisados por 3 meses, obtendo-se curvas sigmoidais. Os esporos aumentaram a partir de 22ºC, atingindo valores maiores em 28ºC e em 32ºC reduziu-se a produção. A maior produção de esporos de G. decipiens ocorreu em pH 6,5, enquanto G. clarum em pH 4,0. O cultivo de outras espécies faz-se necessário visando o conhecimento de aspectos particulares da simbiose até hoje não elucidados, incluindo-se os efeitos dos fatores ambientais sobre a produção dos esporos das diversas espécies de FMAs.

Monoxenic cultures of arbuscular mycorrhizal fungi (AMF) in transformed roots have been used to better understand the symbiosis with these fungi, but few species have been successfully established in vitro. The object was to establish monoxenic cultures of Gigaspora decipiens and Glomus clarum and to verify the effects of temperature and pH on spore formation. Cultures were established from superficially disinfested spores that were germinated on an agar and water. After germination, the spores were transferred to Petri dishes containing transformed carrot roots. After 4-6 days formed newly formed spores and structures typical. The effects of temperature (22, 25, 28 or 32ºC) and pH (4.0, 4.5, 5.5 or 6.5) on the production of spores were assessed over three months, resulting in sigmoidal growth curves. The spore increased from 22ºC, reaching higher values 28oC and 32oC there was a reduction in the production. The highest spore production of G. decipiens occurred in a pH of 6.5, whereas for G. clarum was pH of 4.0. The cultivation of other species is still necessary to elucidate particular aspects of the symbiosis that so far are unclear, including the effects of environmental factors on the production of spores of different AMF species.

In nature, orchid seeds obtain the nutrients necessary for germination by degrading intracellular fungal structures formed after colonization of the embryo by mycorrhizal fungi. Protocols for asymbiotic germination of orchid seeds typically use media with high concentrations of soluble carbohydrate and minerals. However, when reintroduced into the field, seedlings obtained via asymbiotic germination have lower survival rates than do seedlings obtained via symbiotic germination. Tree fern fiber, the ideal substrate for orchid seedling acclimatization, is increasingly scarce. Here, we evaluated seed germination and protocorm development of Cyrtopodium glutiniferum Raddi cultivated in asymbiotic media (Knudson C and Murashige & Skoog) and in oatmeal agar (OA) medium inoculated with the mycorrhizal fungus Epulorhiza sp., using non-inoculated OA medium as a control. We also evaluated the performance of tree fern fiber, pine bark, eucalyptus bark, corncob and sawdust as substrates for the acclimatization of symbiotically propagated plants. We determined germination percentages, protocorm development and growth indices at 35 and 70 days of cultivation. Relative growth rates and the effects of substrates on mycorrhizal formation were calculated after 165 days of cultivation. Germination efficiency and growth indices were best when inoculated OA medium was used. Corncob and pine bark showed the highest percentages of colonized system roots. The OA medium inoculated with Epulorhiza sp. shows potential for C. glutiniferum seedling production. Corncob and pine bark are promising substitutes for tree fern fiber as substrates for the acclimatization of orchid seedlings.

Agronomic biomass yields of forage sorghum BRS 655 presented similar results to other energy crops, producing 9 to 12.6 tons/ha (dry mass) of sorghum straw. The objective of this study was to evaluate the lignocellulosic part of this cultivar in terms of its potential in the different unit processes in the production of cellulosic ethanol, measuring the effects of pretreatment and enzymatic hydrolysis. Three types of pre-treatments for two reaction times were conducted to evaluate the characteristics of the pulp for subsequent saccharification. The pulp pretreated by alkali, and by acid followed by delignification, attained hydrolysis rates of over 90%.

The mushroom Pleurotus ostreatus has nutritional and medicinal characteristics that depend on the growth substrate. In nature, this fungus grows on dead wood, but it can be artificially cultivated on agricultural wastes (coffee husks, eucalyptus sawdust, corncobs and sugar cane bagasse). The degradation of agricultural wastes involves some enzyme complexes made up of oxidative (laccase, manganese peroxidase and lignin peroxidase) and hydrolytic enzymes (cellulases, xylanases and tanases). Understanding how these enzymes work will help to improve the productivity of mushroom cultures and decrease the potential pollution that can be caused by inadequate discharge of the agroindustrial residues. The objective of this work was to assess the activity of the lignocellulolytic enzymes produced by two P. ostreatus strains (PLO 2 and PLO 6). These strains were used to inoculate samples of coffee husks, eucalyptus sawdust or eucalyptus bark add with or without 20 % rice bran. Every five days after substrate inoculation, the enzyme activity and soluble protein concentration were evaluated. The maximum activity of oxidative enzymes was observed at day 10 after inoculation, and the activity of the hydrolytic enzymes increased during the entire period of the experiment. The results show that substrate composition and colonization time influenced the activity of the lignocellulolytic enzymes.

Fungos micorrízicos arbusculares (FMAs) são fungos de solo, biotróficos obrigatórios e formadores da simbiose mutualista mais comum na natureza: a micorriza arbuscular (MA). Essa associação ocorre nas raízes da maioria das plantas terrestres, promovendo melhorias no crescimento, desenvolvimento e aumento na tolerância e, ou, resistência das plantas a vários agentes ambientais adversos. Além disso, os FMAs podem ser utilizados como potenciais agentes de controle biológico de doenças de plantas. Esses fungos produzem ainda glomalina, uma proteína que desempenha papel fundamental na estabilidade do solo e bioestabilização de solos contaminados. As diferentes respostas das plantas a essa simbiose podem ser atribuídas à diversidade funcional das MAs, em função da interação FMA-planta-condições ambientais. O estabelecimento e funcionamento da MA durante as condições de estresse envolvem um complexo processo de reconhecimento e desenvolvimento, concomitantemente às alterações bioquímicas, fisiológicas e moleculares em ambos os simbiontes. Além disso, a colonização micorrízica das raízes tem impacto significativo na expressão de genes de diversas plantas que codificam proteínas presumivelmente envolvidas na tolerância ao estresse. Nesse contexto, considerando que os FMAs são essenciais no estabelecimento e adaptação das plantas em locais perturbados, nesta revisão são abordados os mecanismos fisiológicos e moleculares da associação MA responsáveis por essa adaptação e pela maior tolerância das plantas ao estresse.

Arbuscular mycorrhizal fungi (AMF) are soil fungi, obligate biotrophic fungi and form the most common mutualistic symbiosis in nature: the arbuscular mycorrhiza (AM). This association occurs on the roots of most plants, promoting improvements in plant growth and development and increasing tolerance and/or plant resistance to several adverse environmental agents. In addition, AMF can be a potential biological control agent of plant diseases. These fungi produce glomalin, a protein that plays a key role in soil stability and in the biostabilization of contaminated soils. The different responses of plants to this symbiosis can be assigned to the functional diversity of AM, depending of the interaction between AMF, plants and environmental conditions. The establishment and functioning of AMs under stress conditions involves a complex process of recognition and development, accompanied by physiological, biochemical and molecular changes in both symbionts. In addition, the mycorrhizal colonization of roots has a significant impact on the gene expression of several plants that encode proteins presumably involved in stress tolerance. In this context, since the AMF are essential for the establishment and adaptation of plants on disturbed sites, this review covers the molecular and physiological mechanisms of the AM association, responsible for this adaptation and greater stress tolerance of plants.

Xylanolytic enzymes produced by Lentinula edodes UFV70, cultivated in eucalyptus sawdust/rice bran medium, were stable at 50, 60 and 65ºC for 21 hours, losing only 15-25% activity. Fungus incubation at 50ºC for 12 hours and at 65ºC for 24 hours increased the amount of xylose produced.

A associação micorrízica é advinda da associação simbiótica entre alguns fungos do solos e da maioria das raízes das plantas. O eucalipto possui a capacidade de se associar com dois tipos de micorrizas, a micorriza arbuscular e a ectomicorriza, o que depende muito da sua idade. Este trabalho objetivou avaliar a taxa de colonização por fungos micorrízicos arbusculares (MA) e ectomicorrízicos (ECM) e o número de esporos de fungos micorrízicos arbusculares em plantios comerciais de Eucalyptus grandis e Eucalyptus urophylla, com diferentes idades e manejos, no período de dezembro de 2002 a fevereiro de 2004, na região leste de Minas Gerais, Brasil. Em todas as coletas e em todas as idades dos plantios, foram encontradas MA, e ECM e a média geral da colonização por fungos micorrízicos arbusculares (FMA) foi de 26%. O número médio de esporos desses fungos de 374,7 por 100 g de solo e a colonização por fungos ectomicorrízicos (FECM) de 20,2%. As maiores porcentagens de colonização por FECM foram observadas em áreas de plantios mais jovens, mas a sua maior diversidade foi observada nas áreas de plantios mais velhos. Conclui-se que a época de coleta, a idade do plantio e o manejo do solo afetam a colonização micorrízica e também a diversidade de fungos ECM e que a sucessão de eucalipto com FMA-FECM não é regra e é muito influenciada pelo tipo de manejo.

Mycorrhyzal association is the symbiotic association among some soil fungus and most plant roots. Eucalyptus has the capacity of associating with two types of mychorrizal, arbuscular and ectomycorrhizal mycorrhizal,depending of its age. The objective of this work was to evaluate colonization rates by arbuscular mycorrhizal fungi (AM) and ectomycorrhizal fungi (ECM) and the number of spores of arbuscular mycorrhizal fungi in in commercial plantations of Eucalyptus grandis and Eucalyptus urophylla, at different ages and managements practices, from December 2002 to February, 2004, in eastern region of Minas Gerais, Brazil. In all periods of sampling and at all ages of the plant, it was found both AM and ECM and the colonization general mean by arbuscular mycorrhizal fungi (AMF) was 26%. The average number of spores of those fungi was 374.7 per 100 g of soil and colonization by ectomycorrhizal fungi (EMF) was 20.2 %. Most percentages of colonization by EMF were observed in areas with younger plantations, but it greatest diversity was observed in older plantation areas. It can be concluded that sampling period, age of the plantation and soil management practices affect mycorrhizal colonization and also ECM fungi diversity and succession of eucalyptus with AM-ECM is not a rule and it is strongly affected by management.